LOCUS       sequence-214591-        7458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentivirally express gRNA targeting human TAF12 HFD with GFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7458)
  AUTHORS   Xu Y, Milazzo JP, Somerville TDD, Tarumoto Y, Huang YH, Ostrander 
            EL, Wilkinson JE, Challen GA, Vakoc CR
  TITLE     A TFIID-SAGA Perturbation that Targets MYB and Suppresses Acute 
            Myeloid Leukemia.
  JOURNAL   Cancer Cell. 2018 Jan 8;33(1):13-28.e8. doi: 
            10.1016/j.ccell.2017.12.002.
  PUBMED    29316427
REFERENCE   2  (bases 1 to 7458)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2017.12"; journalName: "Cancer Cell"; date: 
            "2018-01-8- 8"; volume: "33"; issue: "1"; pages: "13-28.e8"
COMMENT     Sequence Label: LRG-2.1T-sgTAF12(human)#4.5
FEATURES             Location/Qualifiers
     source          1..7458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..32
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    149..170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(185..190,191..208,209..215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    223..239
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     228..250
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     247..263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     247..263
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     282..302
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        284..302
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        330..556
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             557..737
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    784..909
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1402..1635
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1820..1864
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2042..2290
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2042..2062
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2213..2232
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2455..2572
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2641..2852
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2872..2881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2881..2904
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2905..2907,2908..2910,2911..3624)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2905..2907,2908..2910,2911..3621)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2950..2971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3211..3230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3558..3579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3640..4228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3693..3713)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4111..4122)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4300..4533
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4611..4732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4648..4667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4702..4721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4772..4907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4927..4946)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4928..4946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4956..4973)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4956..4972)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4965..4987)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5114..5569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5201..5220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5411..5432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5595..5699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5700..5768,5769..6560)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5918..5937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6731..7319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7220..7239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
       61 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
      121 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      181 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      241 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
      301 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
      361 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
      421 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
      481 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
      541 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      601 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
      661 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
      721 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
      781 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
      841 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
      901 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
      961 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     1021 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     1081 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     1141 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     1201 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     1261 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     1321 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     1381 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     1441 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     1501 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     1561 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     1621 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     1681 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     1741 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     1801 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     1861 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     1921 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     1981 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccag
     2041 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     2101 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     2161 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     2221 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     2281 acgaaacacc gctggacatc tttcacctcc gtttaagagc tatgctggaa acagcatagc
     2341 aagtttaaat aaggctagtc cgttatcaac tttgctggaa acagcaaagt ggcaccgagt
     2401 cggtgctttt tttgaattct agatcttgag acaaatggca gtattcatcc acaattttaa
     2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     2581 ttacagggac agcagagatc cactttggcg ccggctcgag tggctccggt gcccgtcagt
     2641 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2701 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2761 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2821 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg
     2881 gattacaaag acgatgacga taagatggtg agcaagggcg aggagctgtt caccggggtg
     2941 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3001 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3061 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3121 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3181 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3241 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3301 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3361 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3421 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3481 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3541 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3601 ggcatggacg agctgtacaa gtagacgcgt taagtcgaca atcaacctct ggattacaaa
     3661 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3721 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3781 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3841 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3901 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3961 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4021 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     4081 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4141 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4201 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
     4261 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     4321 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     4381 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     4441 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     4501 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
     4561 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     4621 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4681 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4741 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4801 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4861 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     4921 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     4981 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5041 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5101 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5161 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5221 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5281 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5341 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5401 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5461 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5521 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     5581 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5881 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5941 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6601 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6841 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7321 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7381 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7441 ataccgctcg ccgcagcc
//
